Imaging subcellular

Kottke, Electron energy loss spectroscopy and imaging techniques for subcellular localization of elements in mycorrhizas. Methods Mierohiol. 23 369 (1991). 

An assay that produces multiple biological readouts. Most commonly used in relation to the mathematical analysis of an image acquired using an automated microscope whereby analysis algorithms quantify cellular parameters (e.g., number, motility, neurite outgrowth, size, shape) and subcellular events (e.g., receptor internalization, protein translocation, protein expression nuclei shape). 

Zelazny, E., Borst, J. W., Muylaert, M., Batoko, H., Hemminga, M. A. and Chaumont, F. (2007). FRET imaging in living maize cells reveals that plasma membrane aquaporins interact to regulate their subcellular localization. Proc. Natl. Acad. Sci. USA 104, 12359-64. 

Coulon, V., Audet, M., Homburger, V., Bockaert, J., Fagni, L., Bouvier, M. and Perroy, J. (2008). Subcellular imaging of dynamic protein interactions by bioluminescence resonance energy transfer. Biophys. J. 94,1001-9. 

A combinatorial library of fluorescent NBD molecules was used to visualize subcellular transport pathways in living cells, using a kinetic, high content imaging system to monitor spatiotemporal variations of intracellular probe distribution <2004MI414>. 

Epi-illumination Subcellular imaging structures Freeze fracture Preparation of cellular ultrastructures in frozen-hydrated and living state for electron microscopy macromolecular organization of bilayer membranes... 

Phase contrast cell boundaries and cell walls also images fibrous subcellular components, e.g., microtubules. Produces visible differences Transmission Electron Subcellular morphology... 

In contrast to many chemotherapeutic agents in cancer therapy, boron compounds for BNCT do not require a tumoricidal action in their own right. For their successful application in the therapy of patients, it is important to deliver, to the tumor, a radiation dose which is higher than the radiation dose to the surrounding tissue. The demonstration that this is actually achieved lies ultimately in the treatment of the tumor in question. Because of the short range of the particles produced in the 10B(n,a)7Li reaction, it is very important where, on a cellular and subcellular dimension, the neutron capture reaction takes place. Different methods for boron detection and quantification give different resolution of the boron distribution. It is instructive to compare these methods, both for their precision and lower detection limits, as well as for their ability to yield an image of the boron distribution in tissue (Table 2.2-1). 

Bader AN, Hofrnan EG, Voortman J, en Flenegouwen PM, Gerritsen HC (2009) Homo-ERET imaging enables quantification of protein cluster sizes with subcellular resolution. Biophys J 97 2613-22... 

With the help of a micro-Raman setup the laser spot can be focused down to about 1 pm in diameter. This allows for the differentiation of single bacterial cells or a biochemical analysis of subcellular components within bacterial (diameter approx. 1 pm) or yeast cells (diameter approx. 5-10pm). A confocal Raman setup achieves an even better spatial resolution [6, 7]. This possibility enables Raman mapping or imaging experiments with spatially resolved information of the whole sample in axial and lateral directions. 

Altelaar, A. F., van Minnen, J, Jimenez, C. R., Heeren, R. M., and Piersma, S. R. (2005). Direct molecular imaging of Lymnaea stagnalis nervous tissue at subcellular spatial resolution by mass spectrometry. Anal. Chem. 77 735-741. 

Here we describe the construction and use of adenoviruses to express lucif-erase- or GFP-fusion proteins for high-throughput cell-based screens. We constructed adenoviruses that express a cyclin Bl-luciferase reporter protein or unmodified luciferase. This enabled us to screen for small molecules that specifically up-regulated levels of the cyclin B 1-luciferase fusion protein without affecting luciferase. A similar approach was used to construct adenoviruses that express cyclin A-GFP or cyclin Bl-GFP fusion proteins. These viruses were used in an imaging-based screen to identify small molecules that specifically upregulated these proteins or altered their subcellular localization. 

R.V. Moore, and A.C. Bovik. 2002. Direct Raman imaging techniques for study of the subcellular distribution of a dmg. 

McDonnell L, Piersma S, Altelaar A, Mize T, Luxembourg S, Verhaert P, van Minnen J, Heeren R (2005) Subcellular imaging mass spectrometry of brain tissue. J Mass Spectrom 40 160-168. doi 10.1002/jms.735... 

The subcellular distribution of the 5-HT receptors likely contributes to their distinct functions. In polarized cells, several lines of evidence support the notion that receptor interaction with FRAPs targets and anchors 5-HT receptors to the appropriate subcellular compartments and modulates receptor internalization. We should also keep in mind that although most studies described in this section capture only a static image, we have only recently begun to appreciate the... 

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